文献类型：期刊文献

英文题名：Construction and characterization of the recombinant immunotoxin RTA-4D5-KDEL targeting HER2/neu-positive cancer cells and locating the endoplasmic reticulum

作者：Jiao, Pengyan[1];Zhang, Jian[1];Dong, Yuguo[1];Wei, Dongzhi[1];Ren, Yuhong[1]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, New World Inst Biotechnol, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2018

卷号：102

期号：22

起止页码：9585

外文期刊名：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY

收录：;EI(收录号：20183505751970);WOS:【SCI-EXPANDED(收录号:WOS:000448593800014)】；

基金：This work was funded by the National Natural Science Foundation of China (No. 21778018 and No. 21706072) and the National Special Fund for the State Key Laboratory of Bioreactor Engineering (2060204).

语种：英文

外文关键词：Recombinant immunotoxin; Ricin A; HER2; Localization peptide; Specific targeting; Cancer cell

摘要：The specific targeting of immunotoxins enables their wide application in cancer therapy. The A-chain of the ricin protein (RTA) is an N-glycosidase that catalyzes the removal of adenine from the 28S rRNA, preventing protein translation and leading to cell death. Ricin is highly toxic but can only exert its toxic effects from within the cytoplasm. In this study, we linked the anti-HER2 single-chain variable fragment 4D5 scFv and the endoplasmic reticulum-targeting peptide KDEL to the C-terminal of the RTA to construct immunotoxin RTA-4D5-KDEL. In vitro experiments showed that the anticancer effect of RTA-4D5-KDEL towards ovarian cancer cells SKOV-3 increased 440-fold and 28-fold relative to RTA and RTA-4D5, respectively. RTA-4D5-KDEL had a strong inhibitory effect on HER2-overexpressing SKOV-3 cells and caused little damage to normal HEK-293 cells and H460 lung cancer cells. Immunofluorescence experiments showed that the immunotoxin RTA-4D5 could specifically bind to SKOV-3 cells, but not to normal cells HEK-293. The immunotoxin RTA-4D5-KDEL could rapidly localize the recombinant protein to the endoplasmic reticulum. These results suggest that the recombinant immunotoxin RTA-4D5-KDEL has a strong inhibitory effect on ovarian cancer cells that overexpress HER2 but little harm to the normal cells.